EP3778619A1 - Segment für oligonukleotidsynthese, herstellungsverfahren dafür und oligonukleotidsyntheseverfahren unter verwendung desselben - Google Patents

Segment für oligonukleotidsynthese, herstellungsverfahren dafür und oligonukleotidsyntheseverfahren unter verwendung desselben Download PDF

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EP3778619A1
EP3778619A1 EP19796884.5A EP19796884A EP3778619A1 EP 3778619 A1 EP3778619 A1 EP 3778619A1 EP 19796884 A EP19796884 A EP 19796884A EP 3778619 A1 EP3778619 A1 EP 3778619A1
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group
protecting group
oligonucleotide
synthesis
formula
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English (en)
French (fr)
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EP3778619A4 (de
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Masanori Kataoka
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Natias Inc
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Natias Inc
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H1/00—Processes for the preparation of sugar derivatives
    • C07H1/02—Phosphorylation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • C07H21/02—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with ribosyl as saccharide radical
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • C07H21/04—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

  • the present invention relates to a segment for use in synthesis of an oligonucleotide, a method for producing the same, and a method for synthesizing an oligonucleotide using the same.
  • nucleic acid drugs having a natural or non-natural oligonucleotide as basic skeleton In recent years, attention has been focused on nucleic acid drugs having a natural or non-natural oligonucleotide as basic skeleton. Production methods by chemical synthesis are widely used to obtain nucleic acid drugs designed to obtain an intended effect.
  • Non-Patent Literature 1 In oligonucleotide synthesis by conventional methods, using a monomer amidite as raw material, a method of extending the length of a nucleotide through a coupling reaction for each base step by step is mainly used (refer to Non-Patent Literature 1).
  • the extension of bases one by one in the oligonucleotide synthesis requires a coupling reaction between a monomer amidite and the 5'-hydroxyl group of a nucleoside, and in addition to that, a step of oxidizing or sulfurizing phosphite and a step of deprotecting the protecting group for the 5'-hydroxyl group of the nucleoside in preparation for a subsequent coupling reaction.
  • Examples of the typical by-products generated in the method for synthesizing an oligonucleotide N-mer by extending bases one by one include an (N-1)-mer which is shorter by one base and an (N-2)-mer which is shorter by two bases, generated in coupling steps.
  • Such (N-1)-mer and (N-2)-mer are very similar in structure and physical properties to the target N-mer.
  • a solid-phase synthesis method including extending chain in the 5'-direction of a nucleoside with a 3'-terminal fixed to a solid-support is employed.
  • a commercially available monomer amidite for use in solid-phase synthesis is required to be purchased or a monomer amidite is required to be self-prepared by an experimenter.
  • commercially available monomer amidites are expensive, obstructing a large amount of target oligonucleotides to be synthesized.
  • a target monomer amidite at high purity is required to be prepared at high yield. The reason is that use of a not high-purity monomer nucleotide has a negative effect on the coupling reaction in oligonucleotide synthesis with high probability.
  • the segment for use in synthesis of an oligonucleotide, a method for producing the same, and a method for synthesizing an oligonucleotide using the same according to the present invention employ the following means.
  • a first aspect of the present invention relates to a segment for use in synthesis of an oligonucleotide, represented by the following formula (I).
  • B is independently a protected or unprotected nucleoside base;
  • R 1 is a protecting group;
  • R 5 is a substituted or unsubstituted aliphatic group, or a substituted or unsubstituted aromatic group;
  • Y is independently H, NHR 6 , a halogen, CN, CF 3 or a hydroxyl group protected with an acyl protecting group, an ether protecting group or a silyl protecting group;
  • R 6 is H, an aliphatic group or an aromatic group;
  • Z is independently H, an alkyl, an O-alkyl, an N-alkyl or a halogen, or forms a Z-Y bond with Y; and (m+n) is an integer of
  • the protecting group may be an acyl protecting group.
  • R 1 may be a protecting group removable under acidic conditions or a trialkylsilyl group; Y may be preferably H or a hydroxyl group protected with a t-butyldimethylsilyl group; Z may be preferably H; and R 5 may be an isopropyl group.
  • a second aspect of the present invention relates to a method for producing a segment for use in synthesis of an oligonucleotide, represented by the following formula (I).
  • B is independently a protected or unprotected nucleoside base;
  • R 1 is a protecting group;
  • R 5 is a substituted or unsubstituted aliphatic group, or a substituted or unsubstituted aromatic group;
  • Y is independently H, NHR 6 , a halogen, CN, CF 3 or a hydroxyl group protected with an acyl protecting group, an ether protecting group or a silyl protecting group;
  • R 6 is H, an aliphatic group or an aromatic group;
  • Z is independently H, an alkyl, an O-alkyl, an N-alkyl or a halogen, or forms a Z-Y bond with Y; and (m+n) is an integer of
  • the production method comprises:
  • the protecting group may be an acyl protecting group.
  • R 1 may be a protecting group removable under acidic conditions or a trialkylsilyl group; Y may be preferably H or a hydroxyl group protected with a t-butyldimethylsilyl group; Z may be preferably H, and R 5 may be an isopropyl group.
  • a step of reacting the compound represented by formula (IV) obtained in the step (a) or the compound represented by formula (VII) obtained in the step (b) with an oxidizing agent or sulfurizing agent may be included on an as needed basis.
  • a third aspect of the present invention relates to a method for synthesizing an oligonucleotide using a segment for use in synthesis of an oligonucleotide, represented by formula (I) .
  • the synthesis method includes (a) a condensation step of condensing an amidite moiety of the segment for use in synthesis of an oligonucleotide represented by formula (I) with a hydroxyl group of a nucleoside or nucleotide, (b) an oxidation step of oxidizing a phosphite-linkage position obtained in the condensation step, and (c) a deprotection step of deprotecting the terminal protecting group of the segment for use in synthesis of an oligonucleotide condensed with a nucleoside or nucleotide in the condensation step.
  • each of the steps may be performed in a solution.
  • each of the steps may be performed on a solid-support.
  • the number of steps can be reduced.
  • the yield of the oligonucleotide having a target length can be therefore improved than that of a conventional method.
  • the segment for use in synthesis of an oligonucleotide in the present embodiment has a structure represented by the following formula (I):
  • B is independently a protected or unprotected nucleoside base;
  • R 1 is a protecting group;
  • R 5 is a substituted or unsubstituted aliphatic group, or a substituted or unsubstituted aromatic group;
  • Y is independently H, NHR 6 , a halogen, CN, CF 3 or a hydroxyl group protected with an acyl protecting group, an ether protecting group or a silyl protecting group;
  • R 6 is H, an aliphatic group or an aromatic group;
  • Z is independently H, an alkyl, an O-alkyl, an N-alkyl or a halogen, or forms a Z-Y bond with Y; and (m+n) is an integer of
  • the segment for use in synthesis of an oligonucleotide in the present embodiment is obtained by the following successive steps (1) and (2).
  • An (n+1)mer nucleotide phosphoramidite is synthesized by repeating the step (2) as many times as required (n times).
  • nucleotide dimer or trimer is synthesized by a conventional method, it is necessary to perform a reaction for protecting the 3'-hydroxxyl group of a 5'-protected/3'-unprotected nucleoside, and then perform a reaction for removing the protecting group for 5'-hydroxyl group only, so that a nucleoside having the 3'-hydroxyl group and, on an as needed basis, a nucleoside base moiety, protected with a protecting group and the 5'-hydroxyl group unprotected (hereinafter referred to as "5'-unprotected/3'-protected nucleoside”) is prepared in advance.
  • a nucleoside having both of the 5'-hydroxyl group and the 3'-hydroxyl group unprotected, with one protecting group less than those of a conventional method can be used for synthesis of the segment.
  • the 3', 5'-unprotected nucleoside as one of the main raw materials in synthesis of the segment can be therefore prepared in a smaller number of steps compared with a conventional method, so that a larger amount can be synthesized at low cost in a short time.
  • the nucleoside base in the present embodiment includes a natural base such as an adenyl group, a guanyl group, a cytosinyl group, a thyminyl group and an uracil group, and a modified base such as a 5-methylcytosinyl group, a 5-fluorouracil group, a 7-methylguanyl group and a 7-deazaadenyl group.
  • the amino group in these nucleoside bases includes a benzyl protecting group, an allyl protecting group, a carbamate protecting group and an acyl protecting group.
  • an acyl protecting group such as an acetyl group, a benzoyl group, a phenoxyacetyl group, and an isopropylcarbonyl group is used.
  • the aliphatic group in the present embodiment includes a saturated or unsaturated, linear or branched C 1 -C 18 hydrocarbon, and a saturated or unsaturated cyclic C 3 -C 18 hydrocarbon.
  • a saturated or unsaturated C 1 -C 8 hydrocarbon or a cyclic C 3 -C 8 hydrocarbon is preferred.
  • the aromatic group in the present embodiment includes a carbocyclic aromatic ring such as a phenyl group, and a carbocyclic aromatic ring condensed with a carbocyclic aromatic ring or a non-carbocyclic aromatic ring such as a naphthyl group.
  • the aliphatic group and the aromatic group in the present embodiment may be substituted with a substituent such as a saturated or unsaturated C 1 -C 8 hydrocarbon or C 3 -C 8 cyclic hydrocarbon, a halogen, a cyano group, a nitro group, and an aromatic ring.
  • a substituent such as a saturated or unsaturated C 1 -C 8 hydrocarbon or C 3 -C 8 cyclic hydrocarbon, a halogen, a cyano group, a nitro group, and an aromatic ring.
  • the protecting groups for 5'-, 3'- or 2'-hydroxyl group in the present embodiment include a protecting group removable under acidic conditions, an acyl protecting group, and a silyl protecting group.
  • the protecting groups removable under acidic conditions include an ether protecting group including a substituted or unsubstituted trityl group, a pixyl group, and a substituted or unsubstituted tetrahydropyranyl (THP) group, and 4,4'-dimethoxytrityl group is a typical protecting group.
  • the silyl protecting groups include a trimethylsilyl group, a triethylsilyl group, a t-butyldimethylsilyl group, a t-butyldiphenylsilyl group, and a triphenylsilyl group.
  • the acyl protecting groups include an acetyl group and a benzoyl group.
  • a nucleoside with a crosslink bond between the 5'-position and the 2'-position may be used as a raw material.
  • a bond of (5'-position)-L-O-(2'-position) can be formed, and examples of L include a C 1 -C 6 alkylene group, wherein the intermediate carbon atom may be substituted with an oxygen atom or a nitrogen atom to which an alkyl group is bonded.
  • step (1) after reacting the 3'-hydroxyl group of the 5'-protected/3'-unprotected nucleoside with a phosphitylating agent to produce 3'-phosphoramidite in situ, an activator that activates the amidite moiety is added to the resulting 3'-phosphoramidite without isolation and purification to cause a reaction with 3', 5'-unprotected nucleoside, so that an extension by one base unit is achieved. Further, through a reaction with a phosphitylating agent, 3'-phosphoramidite is produced.
  • activator examples include 1 H -tetrazole, S-ethylthio-1 H -tetrazole, dicyanoimidazole, and a salt of sulfonic acid and azole or tertiary amine, though not limited thereto.
  • the reaction is performed in a dried solvent such as dichloromethane, acetonitrile, tetrahydrofuran, DMF and toluene.
  • step (1) to a solution of 5'-protected/3'-unprotected nucleoside (0.2 to 0.4 M), a phosphitylating agent (1.05 to 1.2 equivalents of 5'-protected/3'-unprotected nucleoside) and an activator (0.4 to 0.7 equivalents of 5'-protected/3'-unprotected nucleoside) are added and stirred at room temperature for 2 to 5 hours. The resulting 3'-phosphoramidite is purified on silica gel, and then a subsequent step (2) is performed.
  • step (2) to the 3'-phosphoramidite obtained in the step (1), 3', 5'-unprotected nucleoside (1.3 to 2.0 equivalents of 5'-protected/3'-unprotected nucleoside) and an activator (2 to 3 equivalents of 5'-protected/3'-unprotected nucleoside) are added to cause a reaction at room temperature for 0.5 to 1.5 hours, and through silica gel filtration and concentration of the filtrate, a nucleotide intermediate having an unprotected hydroxyl group at a 3'-terminal, with extension by one base unit, is obtained.
  • the yield is about 60 to 95%.
  • the solution of the nucleotide intermediate (0.05 to 0.4 M) is reacted with a phosphitylating agent (1.2 to 2.0 equivalents of 5'-protected/3'-unprotected nucleoside) and an activator (0.5 to 1.0 equivalent of 5'-protected/3'-unprotected nucleotide) to obtain 3'-phophoroamidite of a trimer nucleotide.
  • a phosphitylating agent 1.2 to 2.0 equivalents of 5'-protected/3'-unprotected nucleoside
  • an activator 0.5 to 1.0 equivalent of 5'-protected/3'-unprotected nucleotide
  • the step (2) is further performed to obtain a tetramer or more nucleotide.
  • segments already being a dimer or more may be condensed to each other to achieve extension by two or more base units at a time.
  • Synthesis of an oligonucleotide using the segment for use in synthesis of oligonucleotide represented by formula (I) may be performed in a solution (hereinafter referred to as “liquid-phase synthesis method”), or may be performed on a solid-support (hereinafter referred to as “solid-phase synthesis method”).
  • the nucleoside at the 3' end of the nucleoside or nucleotide having a 3'-hydroxyl group to which a silyl protecting group or an aliphatic-containing protecting group introduced to increase the solubility in the reaction solvent is used to be subjected to repetition of a condensation step (a) of condensation with a segment for use in synthesis of an oligonucleotide represented by formula (I), an oxidation or sulfurization step (b) of oxidizing or sulfurizing the phosphite-linkage position, and a deprotection step (c) of deprotecting the terminal protecting group of the segment for synthesis of an oligonucleotide condensed with nucleoside or nucleotide in the condensation step.
  • a target oligonucleotide can be obtained through a subsequent treatment under basic conditions.
  • a reaction for activating the 3'-terminal amidite of the segment for use in synthesis of an oligonucleotide represented by formula (I) with an activator so as to be condensed with a nucleoside or nucleotide having 5'- unprotected hydroxyl group is performed.
  • phosphite activator a commonly used phosphite activator may be used, and examples thereof include 1 H -tetrazole, S-ethylthio-1 H -tetrazole, dicyanoimidazole, and a salt of sulfonic acid and azole or a tertiary amine, though not limited thereto.
  • the time required for the condensation reaction is generally about 1 minute to 30 minutes, depending on the scale of the reaction.
  • the intermediate obtained in the condensation step is reacted with an oxidizing agent or a sulfurizing agent to obtain a pentavalent phosphate or a thiophosphate nucleotide.
  • the intermediate obtained in the oxidation or sulfurization reaction is reacted with an anhydrous acidic solution to obtain a nucleotide with 5'-hydroxyl group unprotected.
  • a protecting group for the nucleoside base, a protecting group for the 5'-, 3'- or 2'-hydroxyl group, and a protecting group for phosphoric acid in the phosphate bond are deprotected under deprotection conditions corresponding to the protecting group used.
  • Example 1 a tetramer phosphoramidite which is one of the segments for use in synthesis of an oligonucleotide represented by formula (I) was synthesized. Also, according to the procedure shown in Example 3, a pentamer phosphoramidite which is one of the segments for use in synthesis of an oligonucleotide represented by formula (I) was synthesized. Further, according to the procedure shown in Example 4, an oligonucleotide 18-mer was synthesized using a trimer phosphoramidite which is one example of the compounds represented by formula (I).
  • Step 2 Synthesis of DMTr-T p(OCH2CH2CN) T p(OCH2CH2CN) T (OCH2CH2CN) (N( i -C3H7)2) (2)
  • a dichloromethane solution (23 mL) of the phosphoramidite 1 (10.0 g, 9.21 mmol) was added dropwise to a DMF solution (30 mL) of thymidine (2.90 g, 12.0 mmol) and 1H-tetrazole (1.94 g, 27.6 mmol) and stirred at room temperature for 1 hour.
  • Dichloromethane (46 mL) was added to the reaction solution, and the mixture was then applied to 340 g of silica gel for washing with dichloromethane.
  • Step 3 Synthesis of DMTr-T p(OCH2CH2CN) T p(OCH2CH2CN) T p(OCH2CH2CN) T p(OCH2CH2CN) (N( i- C3H7)2) (3)
  • a dichloromethane solution (70 mL) of the phosphoramidite 2 (10.0 g, 7.01 mmol) was added to a DMF solution (90 mL) of thymidine (2.21 g, 9.11 mmol) and 1H-tetrazole (1.47 g, 21.2 mmol) and stirred at room temperature for 1 hour. After adding dichloromethane (140 mL) to the reaction solution, the mixture was applied to 340 g of silica gel for washing with dichloromethane.
  • reaction solution was added to a DMF solution (90 mL) of N 4 -benzoylcytidine (12.9 g, 39.0 mmol) and 1 H -tetrazole (6.31 g, 90.0 mmol), and stirred at room temperature for 1 hour.
  • a phosphitylating agent NCCH 2 CH 2 OP[N( i -C 3 H 7 ) 2 ] 2 (12.4 mL, 39.0 mmol) and 1H-tetrazole (1.05 g, 15.0 mmol) were added in this order at room temperature.
  • reaction solution was applied to 1500 g of silica gel for washing with ethyl acetate. Further, purification by silica gel column chromatography using dichloromethane-IPA as an elution solvent was performed to obtain a target dimer 5 (22.1 g, yield: 87.3%). MS: 723.2 (MNa + )
  • Step 6 Synthesis of DMTr-T p(OCH2CH2CN) C (OCH2CH2CN) G p(OCH2CH2CN) T (OCH2CH2CN) (N( i -C3H7)2) (6)
  • the reaction solution was added dropwise to distilled water (500 mL) and extracted with dichloromethane (500 mL).
  • the resulting crude product was purified by silica gel column chromatography using hexane-ethyl acetate as an elution solvent to obtain a target 3'-protected thymidine 9 (13 g, yield: 88%).
  • the resulting crude product was treated with a dichloromethane solution (20 mL) of 3% dichloroacetic acid at room temperature for 15 minutes, and then the reaction solution was purified by silica gel column chromatography using dichloromethane-methanol as an elution solvent to obtain a target 5'-unprotected dAT dimer 10 (0.65 g, yield: 58%).
  • the resulting crude product was purified by silica gel column chromatography using dichloromethane-methanol as an elution solvent to obtain a target 5', 3'-protected dAATAT pentamer 13 (0.67 g, yield: 94%) .
  • an oligonucleotide (dT 18-mer) was synthesized from universal linker CPG1000 ⁇ (1.0 ⁇ mol) manufactured by ChemGene as a starting substance by the following protocol.
  • an amidite having a length of a nucleoside trimer or more may be used in synthesis of oligonucleotide. For this reason, in the case where an oligonucleotide having a target length equivalent to N-mer is synthesized, an (N-1)-mer which is shorter by one base, an (N-2)-mer which is shorter by two bases, etc., generated in the coupling step of monomer amidite in general-purpose synthesis of oligonucleotides, are not generated.
  • the segment for use in synthesis of an oligonucleotide in the present embodiment enables to reduce the number of steps required for synthesizing the same N-mer oligonucleotide in comparison with the conventional method of extending one base each at a time.
  • the yield of the oligonucleotide having a target length can be therefore improved.
  • a phosphate bond moiety partially oxidized/sulfurized can be formed at the segment stage. Accordingly, even in the case where only a part of the phosphate bonds in an oligonucleotide is made into an oxidized/sulfurized state different from the phosphate bonds in other parts, an oligonucleotide containing a target modified phosphate bond moiety can be synthesized more easily without changing the synthesis procedure of the oligonucleotide.
  • a nucleoside having 5'-hydroxyl group only protected, or a nucleoside having both of 5'-hydroxyl group and 3'-hydroxyl group unprotected is directly reacted with a phosphitylating agent for amidite preparation. Accordingly, it is not necessary to purchase a commercially available 3'-amidite monomer or prepare the monomer in advance. Therefore, a larger amount of a target segment for use in synthesis of an oligonucleotide can be produced as compared with the conventional method.
  • the segment for use in synthesis of an oligonucleotide in the present embodiment may be used in the case where a large amount of oligonucleotide having a relatively short chain is synthesized by the liquid-phase synthesis method, and also in the case where an oligonucleotide having a long chain is synthesized by the solid-phase synthesis method. Since no by-products having a length of N-1 or N-2 are generated, the burden imposed by purification after synthesis of an N-mer oligonucleotide having a long chain in particular can be reduced, so that a target N-mer oligonucleotide can be obtained through a more convenient purification.

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EP19796884.5A 2018-05-02 2019-05-07 Segment für oligonukleotidsynthese, herstellungsverfahren dafür und oligonukleotidsyntheseverfahren unter verwendung desselben Pending EP3778619A4 (de)

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JP2018088912 2018-05-02
PCT/JP2019/018299 WO2019212061A1 (ja) 2018-05-02 2019-05-07 オリゴヌクレオチド合成用セグメントおよびその製造方法、ならびにそれを用いたオリゴヌクレオチドの合成方法

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EP3778619A1 true EP3778619A1 (de) 2021-02-17
EP3778619A4 EP3778619A4 (de) 2021-06-30

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US (1) US20210269470A1 (de)
EP (1) EP3778619A4 (de)
JP (2) JP7075680B2 (de)
CN (1) CN112424213A (de)
WO (1) WO2019212061A1 (de)

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US20210269470A1 (en) * 2018-05-02 2021-09-02 Natias Inc. Segment for use in synthesis of oligonucleotide, method for producing the same, and method for synthesizing oligonucleotide using the same
US20210317158A1 (en) * 2018-05-02 2021-10-14 Natias Inc. Optically active segment for use in synthesis of stereocontrolled oligonucleotide, method for producing the same, and method for synthesizing stereocontrolled oligonucleotide using the same
JP2022177332A (ja) * 2019-10-24 2022-12-01 日東電工株式会社 オリゴヌクレオチドを製造する方法
EP4089099A1 (de) 2020-01-08 2022-11-16 Nitto Denko Corporation Nukleinsäuresyntheseverfahren unter verwendung von segmentamiditen
WO2022224943A1 (ja) 2021-04-21 2022-10-27 株式会社アマダ 曲げ加工システム、及び分割金型の配置方法
WO2023113038A1 (ja) * 2021-12-17 2023-06-22 リードファーマ株式会社 オリゴヌクレオチドの製造方法
CN114230624A (zh) * 2021-12-22 2022-03-25 上海兆维科技发展有限公司 一种核苷二聚体亚膦酰胺的合成方法

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JPS6270392A (ja) * 1985-09-25 1987-03-31 Nippon Zeon Co Ltd オリゴヌクレオチド化合物の製造法
PT638089E (pt) * 1992-04-15 2001-09-27 Univ Johns Hopkins Sintese de coleccoes uteis e diversas de oligonucleotidos
US5869644A (en) * 1992-04-15 1999-02-09 The Johns Hopkins University Synthesis of diverse and useful collections of oligonucleotidies
US5717085A (en) 1994-11-23 1998-02-10 Terrapin Technologies, Inc. Process for preparing codon amidites
JP3985103B2 (ja) * 1996-08-30 2007-10-03 東亞合成株式会社 新規複合体及びオリゴヌクレオチドの合成方法
JP2006248949A (ja) * 2005-03-09 2006-09-21 Univ Nagoya ヌクレオシド誘導体、ヌクレオチド誘導体及びそれらの製造方法
US7999889B2 (en) 2006-03-29 2011-08-16 Sharp Kabushiki Kaisha Scattering-type display including diffraction reducing layer
DE102009046982A1 (de) 2009-11-23 2011-06-01 Ernst-Moritz-Arndt-Universität Greifswald Festphasengebundene Nukleoside
CN103237805B (zh) 2010-08-23 2016-10-19 皇家学习促进学会/麦吉尔大学 寡核糖核苷酸的嵌段合成
SG11201400135SA (en) * 2011-08-23 2014-03-28 Univ Mcgill Ionic tags for synthesis of oligoribonucleotides
WO2017111137A1 (ja) * 2015-12-22 2017-06-29 味の素株式会社 オリゴヌクレオチドの製造方法
US20210269470A1 (en) * 2018-05-02 2021-09-02 Natias Inc. Segment for use in synthesis of oligonucleotide, method for producing the same, and method for synthesizing oligonucleotide using the same

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US20210269470A1 (en) 2021-09-02
EP3778619A4 (de) 2021-06-30
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